| 499 matches found in 345 documents. Search time: 0.038 seconds. |
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| Title: The refolding , purification , and activity analysis of a rice Bowman-Birk inhibitor expressed in Escherichia coli .
| | Author: Li N Qu LJ Liu Y Li Q Gu H Chen Z | | Journal: Protein Expr . Purif . Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub10024476 Accession (PMID): 10024476 | | Abstract: A putative rice trypsin/chymotrypsin inhibitor of the Bowman-Birk family , RBBI-8 of about 20 kDa , was expressed in Escherichia coli as a fusion protein bearing an N-terminal ( His ) 6 purification tag .
The expressed recombinant protein , rRBBI-8 , is insoluble and accumulates as inclusion bodies .
The insoluble protein was solubilized in 8 M urea under reducing environment and then refolded into its active conformation under optimized redox conditions .
Strategies used to optimize yield and efficiency include selecting the redox system , increasing protein concentration during refolding by adding the denatured protein in a stepwise way , utilizing additives to prevent aggregation , and selecting buffer-exchanging conditions .
A Ni-chelate affinity column was then employed to purify the renatured protein . rRBBI-8 shows strong inhibitory activity against trypsin and it can slightly inhibit chymotrypsin .
In this study , a refolding and purification system was set up for this cysteine-rich recombinant protein expressed in a prokaryotic system . | | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
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| Title: Isolation and characterization of the gene encoding the starch debranching enzyme limit dextrinase from germinating barley .
| | Author: Kristensen M Lok F Planchot V Svendsen I Leah R Svensson B | | Journal: Biochim . Biophys . Acta Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub10350630 Accession (PMID): 10350630 | | Abstract: The gene encoding the starch debranching enzyme limit dextrinase , LD , from barley ( Hordeum vulgare ) , was isolated from a genomic phage library using a barley cDNA clone as probe .
The gene encodes a protein of 904 amino acid residues with a calculated molecular mass of 98 . 6 kDa .
This is in agreement with a value of 105 kDa estimated by SDS-PAGE .
The coding sequence is interrupted by 26 introns varying in length from 93 bp to 825 bp .
The 27 exons vary in length from 53 bp to 197 bp .
Southern blot analysis shows that the limit dextrinase gene is present as a single copy in the barley genome .
Gene expression is high during germination and the steady state transcription level reaches a maximum at day 5 of germination .
The deduced amino acid sequence corresponds to the protein sequence of limit dextrinase purified from germinating malt , as determined by automated N-terminal sequencing of tryptic fragments coupled with matrix assisted laser desorption mass spectrometry .
The sequenced peptide fragments cover 70% of the entire protein sequence , which shows 62% and 77% identity to that of starch debranching enzymes from spinach and rice and 37% identity to Klebsiella pullulanase .
Sequence alignment supports the multidomain architecture and identifies both secondary structure elements of the catalytic ( beta/alpha ) 8-barrel substrate , catalytic residues , and specificity associated motifs characteristic of members of the glycoside hydrolase family 13 which cleave alpha-1 , 6-glucosidic bonds .
A remarkable distribution of the secondary structure elements to individual exons is observed .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
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| Title: Localization of farnesyl diphosphate synthase in chloroplasts .
| | Author: Sanmiya K Ueno O Matsuoka M Yamamoto N | | Journal: Plant Cell Physiol .
Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub10353221 Accession (PMID): 10353221 | | Abstract: The subcellular localization of plant farnesyl diphosphate synthase ( FPPS ) was examined .
Immunocytochemical staining using anti-FPPS1 antibody followed by electron microscopy showed that FPPS1 was localized to chloroplasts of rice mesophyll cells .
Subcellular fractions from wheat leaves were examined by immunoblot analysis .
FPPS was detected in the chloroplast fraction in wheat , and was protected from proteolysis following trypsin treatment of chloroplasts .
FPPS was also detected in the chloroplast fraction of a dicot plant , tobacco | | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
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| Title: Nitrogen retention and plasma amino acids of adults who consumed isonitrogenous diets containing rice and milk or wheat versus their constituent amino acids .
| | Author: Clark HE Moon W-H Bailey LB Panemangelore M | | Journal: Am .
J Clin . Nutr . Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub1036663 Accession (PMID): 1036663 | | Abstract: Nitrogen retention and concentrations of plasma amino acids were compared when young adults consumed isonitrogenous diets containing proteins ( 3 . 0 g of N from rice plus 3 . 0 g of N from milk or 3 . 0 g of N from rice plus 3 . 0 g of N from wheat flour ) or mixtures of their constituent amino acids .
Diets containing proteins induced greater nitrogen retention than did those containing corresponding amounts of amino acids in crystalline form , and the difference between the combinations of proteins was delineated more sharply .
Concentrations of several amino acids were elevated by substituting crystalline amino acids for proteins , especially in postprandial plasma .
Subjects responded differently to addition of the limiting amino acids , lysine and tryptophan , to the diets containing amino acids instead of rice plus wheat .
Therefore , data obtained by means of combinations of cereals and by mixtures of their constituent amino acids can not always be used interchangeably . | | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
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| Title: Molecular cloning and characterization of a cysteine-rich 16 . 6-kDa prolamin in rice seeds .
| | Author: Mitsukawa N Konishi R Uchiki M Masumura T Tanaka K | | Journal: Biosci . Biotechnol . Biochem . Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub10635550 Accession (PMID): 10635550 | | Abstract: An alcohol-soluble storage protein , a 16 . 6-kDa prolamin found in rice seeds , was purified from both the total protein body and purified type I protein body fractions .
The partial amino acid sequences of three tryptic peptides generated from the purified polypeptide were analyzed .
A part of the 16 . 6-kDa prolamin cDNA was amplified from developing seed mRNA by the reverse transcribed polymerase chain reaction using an oligo ( dT ) primer and a primer which was synthesized based on the partial amino acid sequence .
The amplified product was used to isolate the full-length cDNA clone ( lambda RP16 ) from a developing seed cDNA library .
The cDNA has an open reading frame encoding a hydrophobic polypeptide of 149 amino acids .
The polypeptide was rich in glutamine ( 20 . 0% ) , cysteine ( 10 . 0% ) , and methionine ( 6 . 9% ) .
The cysteine content was higher than those of most other rice storage proteins .
Messenger RNA of the 16 . 6-kDa prolamin was detected in seeds , but not in other aerial it issues . | | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
